Subcellular Protein Labeling by a Spatially Restricted Arylamine N-Acetyltransferase

ACS Chemical Biology
Fleur KleinpenningKimberly M Bonger

Abstract

Mapping proteins at a specific subcellular location is essential to gaining detailed insight on local protein dynamics. We have developed an enzymatic strategy to label proteins on a subcellular level using arylamine N-acetyltransferase (NAT). The NAT enzyme activates an arylhydroxamic acid functionality into a nitrenium ion that reacts fast, covalently, and under neutral conditions with nucleophilic residues of neighboring proteins. The electron density on the aromatic ring proved important for probe activation as strong labeling was only observed with an arylhydroxamic acid bearing an electron donating substituent. We further demonstrate that, using this electron rich arylhydroxamic acid, clear labeling was achieved on a subcellular level in living cells that were transfected with a genetically targeted NAT to the nucleus or the cytosol.

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Citations

Apr 6, 2019·Frontiers in Chemistry·Marcus J C LongYimon Aye
Nov 6, 2020·Current Opinion in Chemical Biology·Mike L W J SmeenkKimberly M Bonger
Aug 10, 2020·Current Opinion in Chemical Biology·Katarzyna Radziwon, Amy M Weeks

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Methods Mentioned

BETA
proteomic profiling
enzymatic proximity
acetylation
transfection
confocal microscopy

Software Mentioned

BioID

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